Small molecules to block measles spreading in the central nervous system

小分子阻止麻疹在中枢神经系统中传播

• 批准号: 9986209
• 负责人: Matteo Porotto
• 金额: $ 49.26万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-15 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9986209
• 关键词: Academic Medical Centers; Affect; Antiviral Agents; Antiviral resistance; Attenuated Live Virus Vaccine; Biological Assay; Bioluminescence; Cell Line; Cell fusion; Cell membrane; Cell surface; Cells; Cellular Immunity; Central Nervous System Infections; Child; Child Mortality; Chimeric Proteins; Complex; Complication; Dependence; Developing Countries; Disadvantaged; Disease; Disease Outbreaks; Encephalitis; Evolution; General Population; Glycoproteins; HIV; Hemagglutinin; Home environment; Human; Immune; Immunity; Immunize; Immunocompromised Host; Impairment; Individual; Infant; Infection; Infectious Encephalitis; Lead; Libraries; Measles; Measles virus; Mediating; Membrane; Membrane Fusion; Modeling; Molecular; Molecular Conformation; Motor Neurons; Mutation; Neuraxis; Neurons; Paramyxovirus; Patients; Peptides; Periodicity; Persons; Population; Process; Receptor Cell; Recombinants; Reporting; Research; Resistance; Risk; SLAM protein; Signal Transduction; South Africa; Structure; Subacute Sclerosing Panencephalitis; Testing; Thermodynamics; Tissues; United States; Universities; Vaccinated; Vaccines; Viral; Viral Proteins; Virus; Virus Diseases; Vulnerable Populations; anti-viral efficacy; attenuated measles virus; base; brain tissue; combat; drug candidate; efficacy study; experimental study; high throughput screening; nectin; preference; premature; pressure; prevent; receptor; receptor binding; screening; small molecule; small molecule inhibitor; small molecule libraries; viral resistance; virus envelope

Measles virus (MV) is a leading cause of child mortality in developing countries despite the availability of a live attenuated vaccine for over 40 years. Severely immune-compromised people are particularly at risk for MV. The most serious manifestations of MV infection, including encephalitis, occur in people with impaired cellular immunity. MV affects the central nervous system (CNS) in up to half of routine cases; with adequate cellular immunity the infection is eradicated, but individuals with impaired cellular immunity are at a disadvantage. In a recent MV outbreak in South Africa several people died of MV CNS infection. We analyzed the viruses from these patients and found that specific intra-host evolution of the MV fusion machinery -- receptor binding protein (H) + fusion protein (F) -- had occurred. Normally, the MV F is synthesized and maintained in a pre-fusion state until it reaches the cell surface, and this pre-fusion state is intrinsically unstable and thermodynamically driven to the post-fusion state in a process that requires a signal from H upon H's interaction with receptor. However in the case of the “CNS-adapted” viruses, a mutation in F allows it to promote fusion with less dependence on interaction of H with the two known MV cellular receptors; this F is activated independently of H or receptor. The CNS isolate F represents an ideal target for identifying small molecules that block the spread of MV in the CNS by destabilizing the F protein, and thereby promoting premature folding of F to its post-fusion state. Such compounds will effectively decrease the amount of pre-fusion F that is available to mediate cell- to-cell fusion, ultimately halting spread of MV in the CNS. We have adapted and validated a cell based bioluminescence based High Throughput Screen (HTS) assay. The Columbia University Medical Center HTS facility library will be screened for small molecules that efficiently block the fusion mediated by the F from the CNS isolate. Aim 1: Primary HTS of small molecule libraries will be performed. Orthogonal screenings in cell lines and human neurons will confirm efficacy against live virus. Aim 2: The mechanism of action of selected small molecules will be assessed using specific functional assays. Ex vivo efficacy studies will assess the antiviral activity of small molecules in relevant tissues. Viral evolution studies under the selective pressure of small molecules will determine the potential for emergence of viral resistance.

麻疹病毒（MV）是发展中国家儿童死亡率的主要原因 现场衰减疫苗已有40多年。严重免疫受损的人特别有风险 MV。 MV感染最严重的表现，包括脑炎，发生在有障碍的人中 细胞免疫。 MV在多达一半的常规病例中影响中枢神经系统（CNS）；和 足够的细胞免疫学被放射性感染，但细胞免疫学受损的个体在 不利。 在最近在南非的一次MV爆发中，有几人死于MV CNS感染。我们分析了 这些患者的病毒发现MV融合机械的特定主宿内演变 - 受体结合蛋白（H） +融合蛋白（F）发生了。通常，MV F是合成的，并且 保持在融合前状态，直到到达细胞表面为止，并且该前融合态本质上是 不稳定且热力学驱动到需要H h信号的过程中的融合后状态 H与受体相互作用。但是，在“ CNS适应”病毒的情况下，F中的突变 允许它促进融合，而对H与两个已知MV细胞的相互作用较少依赖 受体；该F独立于H或受体激活。 中枢神经系统分离株F表示识别阻断MV扩散的小分子的理想目标 在中枢神经系统中，通过破坏F蛋白的稳定，从而将F过早折叠到其后融合后折叠 状态。这样的化合物将有效地减少可用于介导细胞的融合前F量 待办事项融合，最终停止MV在中枢神经系统中的传播。我们已经改编并验证了一个基于单元的 基于生物发光的高吞吐量屏幕（HTS）测定法。哥伦比亚大学医学中心 HTS设施库将进行筛选的小分子，以有效地阻止F的融合 来自中枢神经系统分离株。 AIM 1：将执行小分子库的主要HT。细胞系中的正交筛选， 人神经元将确认针对活病毒的效率。 AIM 2：将使用特定功能评估选定的小分子的作用机理 测定。离体效率研究将评估相关组织中小分子的抗病毒活性。病毒性的 小分子选择性压力下的进化研究将确定出现的潜力 病毒抗性。